The basement membrane (BM) proteins laminins, which consist of alpha, beta and gamma chains, play critical roles in the maintenance of tissue structures. One of laminin alpha chains, alpha3 has two isoforms, the truncated form alpha3A and the full-sized form alpha3B. In contrast to alpha3A laminins, little is known about alpha3B laminins. To show the histological distribution of the laminin alpha3B chain, we prepared alpha3B-specific monoclonal antibodies. Immunohistochemical analysis showed that the alpha3B chain was colocalized with the alpha3A, beta3 and gamma2 chains in the epithelial BMs of the skin, esophagus, breast and lung, suggesting the presence of laminin-3B32 (laminin-5B) and laminin-3A32 (laminin-5A). In the lung alveoli, laminin-3B32 was dominant over laminin-3A32, but vice versa in other epithelial BMs. In contrast, the BMs of blood vessels including capillaries were strongly positive for alpha3B, but almost or completely negative for alpha3A, beta3 and gamma2. alpha3B was colocalized with beta1 and gamma1 in these BMs. The alpha3B chain was scarcely detected in the vessels of malignant skin cancers, though the gamma2 and beta3 chains were highly expressed in the cancer cells. These results strongly suggest that the laminin alpha3B chain is widely expressed in vascular BMs of normal tissues, probably as laminin-3B11/3B21 (laminin-6B/7B).

Objective: The aim of this study was to show the usefulness of examining HER2 status on fluorescence in situ hybridization using cytological samples taken from recurrent/metastatic tumors. Methods: One hundred freshly aspirated or scraped cytological samples were obtained from locoregional recurrences or distant metastases. Fluorescence in situ hybridization assay for HER2 amplification was performed on both these samples and the formalin-fixed, paraffinembedded tissues of the paired primary tumors of breast cancer, and the relationships between various clinico-pathological factors and HER2 amplification of both tumors were examined. Results: A change in HER2 status was observed in nine cases (9%): six cases (6%) underwent a positive-to-negative conversion in HER2 status and three cases (3%) underwent a negative-to-positive conversion in HER2 status. A positive-to-negative conversion of HER2 status was noted in 4 (36%) of 11 'luminal-B' cases. The change in HER2 status in recurrent or metastatic tumor was noted in more cases treated with drug therapy than in those with no drug therapy (P , 0.05; Fisher's exact probability). Although the time to relapse was 3 years or more in three cases showing a negative-to-positive conversion in HER2 status, the time to relapse was less than 3 years in six cases showing a positive-to-negative conversion (P , 0.05; Fisher's exact probability). Conclusions: HER2 examination on fluorescence in situ hybridization using fine-needle aspiration cytology samples of tumors in recurrent/metastatic sites or disseminated tumor cells in effusion is beneficial, particularly when the primary tumor is not suitable for the testing of HER2 status or negative for HER2 amplification, because aspiration using a needle is technically feasible and not as traumatic as biopsy.

Objectives This study aimed to develop reliable biomarkers that improve the ability of bile cytology to diagnose cholangiocarcinoma vs benign biliary lesions. Methods Many studies indicate that microRNAs (miRNAs) are potential candidates for the early diagnosis of cancer. We analyzed the expression of five tumor-associated miRNAs (miR-31-5p, miR-122-5p, miR-378d, miR-182-5p, and miR-92a-3p) in cytology samples using quantitative reverse transcription polymerase chain reaction. We collected 52 surgically resected tissue samples, 84 cytologic specimens from smears (53 cases of cancer and 31 cases of noncancer), and 40 residual sediments after smearing for routine cytology at Hiroshima University Hospital. Results The expression of miR-31-5p, miR-378d, and miR-122-5p was significantly higher in cancer tissues than those in normal tissues, while miR-182-5p expression was lower. The expression of miR-31-5p, miR-378d, miR-182-5p, and miR-92a-3p was significantly higher in detached cell samples from smears of cholangiocarcinoma cases than in those from noncancer cases. Conclusions These results suggest that the analysis of miRNAs in bile cytologic specimens is a promising auxiliary tool for distinguishing cholangiocarcinoma from benign biliary lesions.

Abstract. the aim of this study was to compare the results of immunohistochemistry (ihc) assays evaluated by human examiners with the results evaluated by computerized image analysis, and to compare the computerized image analysis results among three automated ihc assays, namely the BioGenex, dako and Ventana assays. all slides were semiquantitatively evaluated according to the allred score and J-score by human examiners. the images were analyzed using macScOpE version 2.6 for macintosh according to the h-score and the percentage of positive-stained nuclei per area of carcinoma cells (pp) irrespective of the intensity of the stained nuclei. the h-score for the estrogen receptor (Er) was significantly correlated with the Allred score (P<0.0001) and the PP for the ER was significantly correlated with the J-score (P<0.0001), suggesting that the image analysis used in the present study is a useful method for the evaluation of ER status. Several discrepancies were identified between the allred score and h-score and between the pp and J-score due to the positive-stained cytoplasm area of carcinoma cells and/ or the positive-stained nuclei area of non-carcinoma cells, including benign epithelial cells, lymphocytes and stromal cells. accordingly, advances in the algorithm of the digitized analyzing system is necessary.